Alteration of blood clotting and lung damage by protamine are avoided using the heparin and polyphosphate inhibitor UHRA
Alteration of blood clotting and lung damage by protamine are avoided using the heparin and polyphosphate inhibitor UHRA
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DOI：
10.1182/blood-2016-10-747915
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发表时间：
2017-03-09
期刊：
影响因子：
20.3
通讯作者：
Kizhakkedathu, Jayachandran N.
中科院分区：
文献类型：
--
作者：
Kalathottukaren, Manu Thomas;Abraham, Libin;Kizhakkedathu, Jayachandran N.
抗凝治疗相关的出血和病理性血栓形成给住院患者带来严重风险。这两种并发症都可以通过开发新的疗法来缓解，这些疗法可以安全地中和抗凝活性并抑制内在凝血途径的激活剂，例如多磷酸盐（polyP）和细胞外核酸。后一种策略可以减少抗凝剂的使用，从而可能减少出血事件。然而，先前描述的polyP和细胞外核酸的阳离子抑制剂表现出非特异性结合和对血液凝固的不利影响，这限制了它们的使用。事实上，在手术环境中用于抵消肝素相关出血的聚阳离子鱼精蛋白表现出不良反应。为了解决这些临床缺陷，我们开发了一种合成聚阳离子——通用肝素逆转剂（UHRA），它无毒，可以中和肝素的抗凝活性和聚P的促血栓活性。与鱼精蛋白形成鲜明对比的是，我们发现 UHRA 不会与纤维蛋白原相互作用，不会影响凝块形成过程中的纤维蛋白聚合，也不会消除血浆凝固。使用扫描电子显微镜、共聚焦显微镜和凝块溶解测定，我们确认 UHRA 不会掺入凝块中，并且凝块具有正常的纤维蛋白形态，是稳定的。相反，鱼精蛋白与纤维蛋白凝块结合，这可以解释鱼精蛋白如何引发凝块溶解并增加手术后出血。最后，对小鼠的研究表明，UHRA 可逆转肝素抗凝活性，而不会造成鱼精蛋白所见的肺损伤。这里提供的数据表明，在止血的不良治疗调节过程中，UHRA 可以安全地用作解毒剂。 （血。2017；129(10):1368-1379）
Anticoagulant therapy-associated bleeding and pathological thrombosis pose serious risks to hospitalized patients. Both complications could be mitigated by developing new therapeutics that safely neutralize anticoagulant activity and inhibit activators of the intrinsic blood clotting pathway, such as polyphosphate (polyP) and extracellular nucleic acids. The latter strategy could reduce the use of anticoagulants, potentially decreasing bleeding events. However, previously described cationic inhibitors of polyP and extracellular nucleic acids exhibit both nonspecific binding and adverse effects on blood clotting that limit their use. Indeed, the polycation used to counteract heparin- associated bleeding in surgical settings, protamine, exhibits adverse effects. To address these clinical shortcomings, we developed a synthetic polycation, Universal Heparin Reversal Agent (UHRA), which is nontoxic and can neutralize the anticoagulant activity of heparins and the prothrombotic activity of polyP. Sharply contrasting protamine, we show that UHRA does not interact with fibrinogen, affect fibrin polymerization during clot formation, or abrogate plasma clotting. Using scanning electron microscopy, confocal microscopy, and clot lysis assays, we confirm that UHRA does not incorporate into clots, and that clots are stable with normal fibrin morphology. Conversely, protamine binds to the fibrin clot, which could explain how protamine instigates clot lysis and increases bleeding after surgery. Finally, studies in mice reveal that UHRA reverses heparin anticoagulant activity without the lung injury seen with protamine. The data presented here illustrate that UHRA could be safely used as an antidote during adverse therapeutic modulation of hemostasis. (Blood. 2017;129(10):1368-1379)